Garden bean line ex 15340804

ABSTRACT

The invention provides seed and plants of the bean line designated EX 15340804. The invention thus relates to the plants, seeds and tissue cultures of bean line EX 15340804, and to methods for producing a bean plant produced by crossing a plant of bean line EX 15340804 with itself or with another bean plant, such as a plant of another line. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of a plant of bean line EX 15340804, including the pods and gametes of such plants.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding and, more specifically, to the development of bean line EX 15340804.

BACKGROUND OF THE INVENTION

The goal of vegetable breeding is to combine various desirable traits in a single variety/hybrid. Such desirable traits may include greater yield, resistance to insects or pests, tolerance to heat and drought, better agronomic quality, higher nutritional value, growth rate and fruit or grain properties.

Breeding techniques take advantage of a plant's method of pollination. There are two general methods of pollination: a plant self-pollinates if pollen from one flower is transferred to the same or another flower of the same plant or plant variety. A plant cross-pollinates if pollen comes to it from a flower of a different plant variety.

Plants that have been self-pollinated and selected for type over many generations become homozygous at almost all gene loci and produce a uniform population of true breeding progeny, a homozygous plant. A cross between two such homozygous plants produces a uniform population of hybrid plants that are heterozygous for many gene loci. Conversely, a cross of two plants each heterozygous at a number of loci produces a population of hybrid plants that differ genetically and are not uniform. The resulting non-uniformity makes performance unpredictable.

The development of uniform varieties requires the development of homozygous inbred plants, the crossing of these inbred plants, and the evaluation of the crosses. Pedigree breeding and recurrent selection are examples of breeding methods that have been used to develop inbred plants from breeding populations. Those breeding methods combine the genetic backgrounds from two or more plants or various other broad-based sources into breeding pools from which new lines are developed by selfing and selection of desired phenotypes. The new lines are evaluated to determine which of those have commercial potential.

One crop species which has been subject to such breeding programs and is of particular value is the bean (Phaseolus vulgaris). Beans are annual, warm-season legumes. Garden beans (Phaseolus vulgaris (snap)), also known as green beans, snap beans, French beans, wax beans, string beans, haricot vert, or pole beans, are grown primarily for their pods, which are harvested for consumption in their succulent form; whereas dry beans (Phaseolus vulgaris (dry)), lima beans (Phaseolus lunatus), and soybeans (Glycine max) are grown primarily for the seed itself. In addition, the bean leaf is occasionally used as a leaf vegetable, and the straw is used for fodder.

Beans are available in bush and pole varieties. Bush varieties form erect bushes 20-60 centimeters tall, while pole varieties form vines 2-3 meters long. The pods are typically 8-20 centimeters long, 1-1.5 centimeters wide, and either green, yellow, black or purple in color. Each pod generally contains 4-6 beans.

The bean is a diploid (2n=2x=22) plant. Genetic improvement of bean has been achieved largely through the selection of varieties by applying conventional breeding techniques of self-pollinated crops. While breeding efforts to date have provided a number of useful bean lines with beneficial traits, there remains a great need in the art for new lines with further improved traits. Such plants would benefit farmers and consumers alike by improving crop yields and/or quality.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a bean plant of the line designated EX 15340804. Also provided are bean plants having all the physiological and morphological characteristics of the bean line designated EX 15340804. Parts of the bean plant of the present invention are also provided, for example, including pollen, an ovule, a pod, and a cell of the plant.

The invention also concerns seed of bean line EX 15340804. The bean seed of the invention may be provided as an essentially homogeneous population of seed of the line designated EX 15340804. Essentially homogeneous populations of seed are generally free from substantial numbers of other seed. Therefore, seed of line EX 15340804 may be defined as forming at least about 97% of the total seed, including at least about 98%, 99%, or more of the seed. The population of bean seed may be particularly defined as being essentially free from hybrid seed. The seed population may be separately grown to provide an essentially homogeneous population of bean plants designated EX 15340804.

In another aspect of the invention, a plant of bean line EX 15340804 comprising an added heritable trait is provided. The heritable trait may comprise a genetic locus that is a dominant or recessive allele. In one embodiment of the invention, a plant of bean line EX 15340804 is defined as comprising a single locus conversion. In specific embodiments of the invention, an added genetic locus confers one or more traits such as, for example, herbicide tolerance, insect resistance, disease resistance, and modified carbohydrate metabolism. The trait may be, for example, conferred by a naturally occurring gene introduced into the genome of the line by backcrossing, a natural or induced mutation, or a transgene introduced through genetic transformation techniques into the plant or a progenitor of any previous generation thereof. When introduced through transformation, a genetic locus may comprise one or more transgenes integrated at a single chromosomal location.

In another aspect of the invention, a tissue culture of regenerable cells of a plant of line EX 15340804 is provided. The tissue culture will preferably be capable of regenerating plants capable of expressing all of the physiological and morphological characteristics of the line, and of regenerating plants having substantially the same genotype as other plants of the line. Examples of some of the physiological and morphological characteristics of the line EX 15340804 include those traits set forth in the tables herein. The regenerable cells in such tissue cultures may be derived, for example, from embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, seed and stalks. Still further, the present invention provides bean plants regenerated from a tissue culture of the invention, the plants having all the physiological and morphological characteristics of line EX 15340804.

In yet another aspect of the invention, processes are provided for producing bean seeds, plants and pods, which processes generally comprise crossing a first parent bean plant with a second parent bean plant, wherein at least one of the first or second parent bean plants is a plant of the line designated EX 15340804. As used herein, “bean plant” refers to any type of bean sexually compatible with a plant of the invention and specifically including garden and dry bean. It will therefore be understood by those of skill in the art that the aforementioned and other methods described herein involving crossing a plant of the invention with a second bean plant may comprise, in one embodiment, crossing a plant of EX 15340804 with a bean plant categorized as a dry bean or garden bean plant. These processes may be further exemplified as processes for preparing hybrid bean seed or plants, wherein a first bean plant is crossed with a second bean plant of a different, distinct line to provide a hybrid that has, as one of its parents, the bean plant line EX 15340804. In these processes, crossing will result in the production of seed. The seed production occurs regardless of whether the seed is collected or not.

In one embodiment of the invention, the first step in “crossing” comprises planting seeds of a first and second parent bean plant, often in close proximity so that pollination and fertilization will occur for example, mediated by insect vectors. Alternatively, pollen can be transferred manually. Where the plant is self-pollinated, pollination and fertilization may occur without the need for direct human intervention other than plant cultivation.

A second step may comprise cultivating or growing the seeds of first and second parent bean plants into plants that bear flowers. A third step may comprise preventing self-pollination of the plants, such as by emasculating the male portions of flowers, (i.e., treating or manipulating the flowers to produce an emasculated parent bean plant). Self-incompatibility systems may also be used in some hybrid crops for the same purpose. Self-incompatible plants still shed viable pollen and can pollinate plants of other varieties but are incapable of pollinating themselves or other plants of the same line.

A fourth step for a hybrid cross may comprise cross-pollination between the first and second parent bean plants. Yet another step comprises harvesting the seeds from at least one of the parent bean plants. The harvested seed can be grown to produce a bean plant or hybrid bean plant.

The present invention also provides the bean seeds and plants produced by a process that comprises crossing a first parent bean plant with a second parent bean plant, wherein at least one of the first or second parent bean plants is a plant of the line designated EX 15340804. In one embodiment of the invention, bean seed and plants produced by the process are first generation (F₁) hybrid bean seed and plants produced by crossing a plant in accordance with the invention with another, distinct plant. The present invention further contemplates plant parts of such an F₁ hybrid bean plant, and methods of use thereof. Therefore, certain exemplary embodiments of the invention provide an F₁ hybrid bean plant and seed thereof.

In still yet another aspect of the invention, the genetic complement of the bean plant line designated EX 15340804 is provided. The phrase “genetic complement” is used to refer to the aggregate of nucleotide sequences, the expression of which sequences defines the phenotype of, in the present case, a bean plant, or a cell or tissue of that plant. A genetic complement thus represents the genetic makeup of a cell, tissue or plant, and a hybrid genetic complement represents the genetic make up of a hybrid cell, tissue or plant. The invention thus provides bean plant cells that have a genetic complement in accordance with the bean plant cells disclosed herein, and plants, seeds and plants containing such cells.

Plant genetic complements may be assessed by genetic marker profiles, and by the expression of phenotypic traits that are characteristic of the expression of the genetic complement, e.g., isozyme typing profiles. It is understood that line EX 15340804 could be identified by any of the many well known techniques including, but not limited to, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., 1998).

In still yet another aspect, the present invention provides hybrid genetic complements, as represented by bean plant cells, tissues, plants, and seeds, formed by the combination of a haploid genetic complement of a bean plant of the invention with a haploid genetic complement of a second bean plant, preferably, another, distinct bean plant. In another aspect, the present invention provides a bean plant regenerated from a tissue culture that comprises a hybrid genetic complement of this invention.

In still yet another aspect, the invention provides a plant of a Garden Bean line that exhibits root rot resistance as exhibited by line EX 15340804, wherein the expression of the trait is controlled by genetic means for the expression of the trait found in bean line EX 15340804. On a practical basis root rot results from a mix of soil organisms including, but not limited to, Aphanomyes Root and Hypocotyl Rot, Black Root Rot, Fusarium Root Rot, and Pythium Diseases (see, e.g., Compendium of Bean Diseases, second edition, Eds. Howard F. Schwartz, James R. Steadman, Robert Hall, and Robert L Forster, The American Phytopathology Press., 2005). Quantification of root rot resistance is achieved by comparing lines with a known susceptible reaction to new lines in locations where root rot is considered a problem. Comparisons that might be quantified include, but are not limited to, plant biomass, root biomass, yield of immature pods, and dry seed yield. Other comparisons utilized but more difficult to quantify have included plant vigor and scoring of disease severity base on discoloration of root and hypcotyl.

In still yet another aspect, the invention provides a method of determining the genotype of a plant of bean line EX 15340804 comprising detecting in the genome of the plant at least a first polymorphism. The method may, in certain embodiments, comprise detecting a plurality of polymorphisms in the genome of the plant. The method may further comprise storing the results of the step of detecting the plurality of polymorphisms on a computer readable medium. The invention further provides a computer readable medium produced by such a method.

In still yet another aspect, the invention provides a method of producing a bean plant derived from the bean line EX 15340804, the method comprising the steps of: (a) preparing a progeny plant derived from bean line EX 15340804, wherein said preparing comprises crossing a plant of the bean line EX 15340804 with a second bean plant; and (b) crossing the progeny plant with itself or a second plant to produce a seed of a progeny plant of a subsequent generation. In particular embodiments, the method may further comprise (c) growing a progeny plant of a subsequent generation from said seed of a progeny plant of a subsequent generation and crossing the progeny plant of a subsequent generation with itself or a second plant; and repeating the steps for an additional 3-10 generations to produce a bean plant derived from bean line EX 15340804. The plant derived from bean line EX 15340804 may be an inbred line, and the aforementioned step may be defined as comprising sufficient inbreeding to produce the inbred line. In the method, it may be desirable to select particular plants resulting from step (c) for continued crossing according to steps (b) and (c). By selecting plants having one or more desirable traits, a bean plant derived from the bean line EX 15340804 is obtained which possesses some of the desirable traits of bean line EX 15340804 as well as potentially other selected traits.

These and other features and advantages of this invention are described in, or are apparent from, the following detailed description of various exemplary embodiments of the devices and methods according to this invention.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants, seeds and derivatives of bean line EX 15340804. This line shows uniformity and stability within the limits of environmental influence for the traits described hereinafter. Bean line EX 15340804 provides sufficient seed yield. By crossing with a distinct second plant, uniform F1 hybrid progeny can be obtained.

Line EX 15340804 exhibits a number of improved traits including root rot resistance in combination with elite agronomic characteristics. EX 15340804 is a green garden bean for the processor market in the United States developed by backcross and pedigree breeding methods. Selection of progeny involved greenhouse screening and field performance in both root rot and non root rot nurseries. The line is adapted for processor sales in the North Central, Northeast, and Southeast USA. EX 15340804 has resistance to Bacterial Brown Spot (Pseudomonas syringae). The line can be compared to the line designated Hystyle, but is distinct in at least growth and reproductive response as measured by seed yield and growth under short rotation, root rot conditions in Wisconsin, as demonstrated in the performance data herein.

A. Origin and Breeding History of Bean Line EX 15340804

The line EX 15340804 was developed by backcross and pedigree breeding selection methods from an initial hand pollinated cross between line B373 and a complex cross designated (ZAA3#1/RES-GB#1). B373, which is also designated Hercules, is a commercial line developed by Seminis Vegetable Seeds. B373 is a large sieve green bean used in the processor industry. It is a medium green color and has good resistance to bacterial brown spot.

The second parent in the initial cross from which EX 15340804 was developed resulted from the cross ZAA3#1/RES-GB#1. ZAA3#1 was an F4 bulk of several plants selected for resistance to Aphanomyces euteiches based on greenhouse evaluations. The parents for ZAA3#1 were Californian Light Red Kidney and PI 209488. RES-GB#1 resulted from the cross of two F7+1 lines designated R97.31120/R97.31010. Line R97.31120 was a cross of Profit/HMX8962. Line R97.31010 was a cross of Profit/Tempest. Lines R97.31120 and R97.31010 had been selected for performance under short rotation conditions in the Grand Marsh area of Central Wisconsin, but neither line appeared to be commercially viable. Profit and Tempest were initially developed as commercial lines for the processor market, although neither became an established cultivar and they are no longer marketed.

The breeding history of EX 15340804 can be summarized as follows:

TABLE 1 Summary of Development of EX 15340804 Year Season Gen Description 1998 Spring a′/b′ Cross was made of Line: R97.31120, grown under stake number Aph186, and Line: R97.31010 grown under stake number Aph185. The F1 was designated RES-GB#1. 1998 Summer a/b RES-GB#1 was planted under stake number Aph201 and crossed with ZAA3#1, planted under stake number Aph191. RES-GB#1 was used as the male and (4) Aph191/Aph201 crosses made. 1998 Fall A/B Four F1 seeds of ZAA3#1/RES-GB#1 (parent B) were planted under stake number Aph217 in two pots. Thirty seeds in ten pots of B373 (parent A) were planted under stake number Aph222. Eighteen pollinations were completed A/B, resulting in 51 seeds. 1999 Spring Bc1 Fifty-one seeds of B373 // ZAA3#1/RES-GB#1 were planted, one plant per pot under stake number APH.491. Using APH.491 as a pollen source, crosses were made to B373 planted under stake number APH.222 from BC1 plants. 1999 Summer Bc2 BC2 seeds from the cross B373 /// B373 // ZAA3#1/RES-GB#1 were planted and advanced to BC2S1. EX 15340804 traced back to pot number APH.1156. 1999 Fall Bc2S1 Fifteen BC2S1 seeds were planted in Chile during the off season. Six individual single plant selections were made from plot CH00.2282. EX 15340804 traced to plot CH00.2282.06. 2000 Summer Bc2S2 Seed of individual single plant selections from Chile were planted in the root rot nursery in Grand Marsh, Wisconsin and in Filer, Idaho. Based on performance in the root rot nursery at Grand Marsh, seed was harvested in the respective plots at Filer, Idaho. EX 15340804 traced to the Filer, Idaho plot number R00.35272. 2001 Summer Bc2S3 Seed of R00.35272 was planted in the root rot nursery in Grand Marsh, Wisconsin and in Filer, Idaho. Based on performance in the root rot nursery at Grand Marsh, BC2S3 seed was harvested in the respective plots at Filer, Idaho. EX 15340804 traced to the Filer, Idaho plot number R01.8003 2001 Fall Bc2S3 + 1 Evaluation for Aphanomyces euteiches resistance was carried out in greenhouse screening. 2002 Summer Bc2S3 + 1 Evaluation for field resistance to root rot complex including Aphanomyces euteiches and general horticultural characteristics was carried out. 2002 Fall Bc2S3 + 1 Evaluation for Aphanomyces euteiches resistance in greenhouse screening was carried out. 2003 Winter Bc2S3 + 1 Two hundred pots of R01.8003 were increased in Guatemala and harvested individually as a Generation 0. 2003 Summer Gen 1 Generation 1 produced plot number RWV3228. 2004 Summer Performance evaluations were carried out of green pod and seed production characteristics.

Observations during further multiplication and increases in Idaho indicated that EX 15340804 is uniform and stable within commercially acceptable limits. As is true with other garden beans, a very small percentage of variants or other off-types may occur within commercially acceptable limits for almost any characteristic during the course of repeated multiplication.

B. Physiological and Morphological Characteristics of Garden Bean Line EX 15340804

In accordance with one aspect of the present invention, there is provided a plant having the physiological and morphological characteristics of bean line EX 15340804. A description of the physiological and morphological characteristics of bean line EX 15340804 is presented in Table 2.

TABLE 2 Physiological and Morphological Characteristics of Line EX 15340804 Objective Morphological Comparison of Varieties Descriptor 15340804 Hystyle Explanation of descriptors Maturity Maturity-DAP 54 55 Maturity-HU base 50 1196 1225 Days to 50% bloom 41 41 Plant Description Plant Habit 1 1 1 = Determinate 2 = Indeterminate 3 = Indeterminate 4 = Indeterminate Type I Type II Type III Type IV erect stems prostrate climbing Plant Height in cm 55 46 Plant Width in cm 54 52 Pod Position 1 2 1 = low 2 = high 3 = scattered Bush Form 1 1 1 = spherical 2 = Stem 3 = wide 4 = high Machine harvest 1 1 1 = Adapted 2 = Not Adapted Leaf surface 3 2 1 = dull 2 = glossy 3 = intermediate leaf size 2 2 1 = small 2 = medium 3 = large leaf color 2 2 1 = light green 2 = medium green 3 = dark green Anthocyanin pigment flowers 1 1 1 = absent 2 = present stems 1 1 pods 1 1 seeds 1 1 leaves 1 1 petioles 1 1 peduncles 1 1 nodes 1 1 Flower color Standard 7 7 1 = white 2 = cream 3 = pink 4 = lilac Wings 1 2 5 = purple 6 = blue 7 = light green Keel 1 2 Pod Description Pubescence 2 2 1 = none 2 = Sparse 3 = Considerable Spur length 11 13 (mm) Seeds per pods 7 6 Number Cross Section 4 4 1 = Flat 2 = heart 3 = round 4 = figure eight Creaseback 1 1 1 = Present 2 = Absent Fiber 1 1 1 = none 2 = Sparse 3 = Considerable Suture String 2 2 1 = Present 2 = Absent Seed Development 2 1 1 = Slow 2 = Medium 3 = Fast Pod Color Immature fresh pods 2 1 1 = light green 2 = medium green 3 = dark green 4 = yellow Dry pod color 2 2 1 = buckskin 2 = green 3 = salmon Seed Characteristics Shape: Hilum view 2 2 1 = Elliptical 2 = Oval 3 = Round Shape: Cross Section 2 2 1 = Elliptical 2 = Oval 3 = Cordate 4 = Round Shape: Side View 1 1 1 = Oval to 2 = Round 3 = Reniform Oblong Grams/100 seeds 54.5 55.4 Seed coat luster 3 3 1 = Shiny 2 = Dull 3 = Semishiny 4 = Variable Seed Coat 1 1 1 = Monocbrome 2 = Polychrome Primary Color-seed coat 11 11 1 = white, 2 = Yellow, 3 = Buff, 4 = Tan, 5 = Brown, 6 = Pink, 7 = Red, 8 = Purple, 9 = Blue, 10 = Black, 11 = (specify)     green     Secondary Color-seed na na (See Key for Primary Color) coat Seed Coat Pattern 1 1 1 = Solid, 2 = Splashed, 3 = Mottled, 4 = Striped, 5 = Flecked, 6 = Dotted Hilar Ring 1 1 1 = Absent 2 = Present Hilar Ring Color na na (See Key for Primary Color) Sieve Size Distribution 1&2 6.8 9.4 Percent 3 8.4 11.0 4 27.2 34.2 5 55.4 45.0 6 2.2 0.4 Pod dimension Sieve Size measured Length (cm) 12.2 11.6 5 sieve Width (mm) 9.5 9.3 Thickness (mm) 9.6 9.9 Disease Resistance Reaction Race(s) Anthracnose S S Colletotrichum lindemuthianum Bean Rust S S Uromyces appendiculatus Powdery Mildew NT NT Erysiphe polygoni Fusarium Root Rot NT NT Fusarium solani f.sp. phaseoli Aphanomyces Root Rot R I Aphanomyces euteiches Pythium Root Rot NT NT Pythium spp. Rhizoctonia Root Rot NT NT Rhizoctonia solani Aerial Pythium NT NT Pythium ultimum Angular Leaf Spot NT NT Isariopsis griseola Bacterial Wilt NT NT Corynebacterium flaccumfaciens subsp. Flaccumfaciens Bacterial Brown Spot R R Pseudomonas syringae pv. syringae Common Blight NT NT Xanthomonas campestris pv. Phaseoli Halo Blight S S Pseudomonas syringae pv. phaseoli Clover Yellow Vein NT NT CYVV Virus Bean Common Mosaic R R BCMV Bean Yellow Mosaic NT NT BYMV Curly Top Virus R R BCTV S = Susceptible R = Resistant I = Intermediate NT = Not Tested *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.

Line EX 15340804 has been self-pollinated and planted for a number of generations to produce the homozygosity and phenotypic stability to make this line useful in commercial seed production. No variant traits have been observed or are expected for this line. The line, being substantially homozygous, can be reproduced under self-pollinating or sib-pollinating conditions and harvesting the resulting seeds using techniques known to one of skill in the art.

C. Breeding Bean Line EX 15340804

One aspect of the current invention concerns methods for crossing the bean line EX 15340804 with itself or a second plant and the seeds and plants produced by such methods. These methods can be used for propagation of line EX 15340804, or can be used to produce hybrid bean seeds and the plants grown therefrom. Hybrid seeds are produced by crossing line EX 15340804 with second bean parent line.

The development of new varieties using one or more starting varieties is well known in the art. In accordance with the invention, novel varieties may be created by crossing line EX 15340804 followed by multiple generations of breeding according to such well known methods. New varieties may be created by crossing with any second plant. In selecting such a second plant to cross for the purpose of developing novel lines, it may be desired to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) in progeny. Once initial crosses have been made, inbreeding and selection take place to produce new varieties. For development of a uniform line, often five or more generations of selling and selection are involved.

Backcrossing can also be used to improve an inbred plant. Backcrossing transfers a specific desirable trait or traits from one inbred or non-inbred source to an inbred that lacks that trait(s). This can be accomplished, for example, by first crossing a superior inbred (A) (recurrent parent) to a donor inbred (non-recurrent parent), which carries the appropriate locus or loci for the trait in question. The progeny of this cross are then mated back to the superior recurrent parent (A) followed by selection in the resultant progeny for the desired trait to be transferred from the non-recurrent parent. After five or more backcross generations with selection for the desired trait, the progeny are heterozygous for loci controlling the characteristic being transferred, but are like the superior parent for most or almost all other loci. The last backcross generation would be selfed to give pure breeding progeny for the trait being transferred. Desired plants that are homozygous for the desired trait(s) are then selected from this selfed population.

The line of the present invention is particularly well suited for the development of new lines based on the elite nature of the genetic background of the line. In selecting a second plant to cross with EX 15340804 for the purpose of developing novel bean lines, it will typically be preferred to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Examples of desirable characteristics may include seed yield, seed size, seed shape, seed uniformity, pod size, pod shape, pod color, pod uniformity, stringless pods, early maturity, disease resistance, herbicide resistance, seedling vigor, adaptability for soil conditions, adaptability for climate conditions, and uniform plant height.

D. Performance Characteristics

As described above, line EX 15340804 exhibits desirable agronomic traits, including improved root rot resistance. These and other performance characteristics of the line were the subject of an objective analysis of the performance traits of the line relative to other lines. The results of the analysis are presented below.

TABLE 3 Comparison of seed yields for EX 15340804 and Hystyle over 2 Seminis root rot nurseries in Wisconsin Loc Name Rep Plot kg/ha Ave @ Loc CSET 15340804 1 PB06.1 1420 CSET 15340804 2 PB06.2 390 CSET 15340804 3 PB06.3 1610 CSET 15340804 4 PB06.4 1580 1250 NCSSrre 15340804 1 R06.16535 1559 NCSSrre 15340804 2 R06.16610 2601 NCSSrre 15340804 3 R06.16681 2454 2204 1659 Mean CSET Hystyle 1 PB37.1 100 CSET Hystyle 2 PB37.2 70 CSET Hystyle 3 PB37.3 40 CSET Hystyle 4 PB37.4 20  575 NCSSrre Hystyle 1 R06.16532 672 NCSSrre Hystyle 2 R06.16607 1886 NCSSrre Hystyle 3 R06.16686 749 1102 683 Mean

The location described as CSET in Table 3 is a typical irrigated light textured soil in the Grand Marsh area of Central Wisconsin. The field was confirmed to have root rot production issues including Aphanomyces euteiches, based on observations by growers over 30 years. The field designated NCSSrre is a dry land silt loam soil type located on the Seminis Research Station in DeForest, Wis. Observations over the past 6 years have included stunted plants, poor growth, along with diseased hypocotyls and poor root systems. Aphanomyces euteiches has been successfully isolated from this field.

A two sample T test was carried out analyzing differences between Hystyle and EX 15340804 over 2 locations. The analysis confirms significant differences between the lines (Table 4).

TABLE 4 Statistical analysis over 2 locations in Table 1 for EX 15340804 & Hystyle. Two-Sample T Test for Kg by Variety Variety Mean Sample Size S.D. S.E. 15340804 1659.1 7 730.93 276.26 Hystyle 505.29 7 683.04 258.17 Difference 1153.9 Null Hypothesis: Difference = 0 Alternative Hyp. Difference >0 95% CI for Assumption T DF P Difference Equal Variance 3.05 12 0.0050 (330.01, 1977.7) Unequal Variance 3.05 11.9 0.0051 (329.60, 1978.1) Test of Equality of Variance F Num DF Den DF P 1.15 6 6 0.4368 Cases included: 14 Missing cases: 0

Additional analyses were carried out of the differences between EX 15340804 and the line designated Hercules, which is slightly darker and later in maturity than EX 15340804. The results of the analyses are presented in Tables 4 and 5, and confirm the differences between the lines.

TABLE 5 Comparison of seed yields for EX 15340804 and Hercules over 2 Seminis root rot nurseries in Wisconsin. Loc Name Rep Plot kg/ha Ave @ Loc NCSSrre 15340804 1 R06.16535 1559 NCSSrre 15340804 2 R06.16610 2601 NCSSrre 15340804 3 R06.16681 2454 2204 2204 Mean NCSSrre Hercules 1 R06.16537 1076 NCSSrre Hercules 2 R06.16603 1490 NCSSrre Hercules 3 R06.16685 1352 1306 Mean

TABLE 6 Statistical analysis for EX 15340804 vs Hercules in Table 5 Two-Sample T Test for Kg by Variety Variety Mean Sample Size S.D. S.E. 15340804 2204.7 3 563.97 325.61 Hercules 1306.0 3 210.80 121.70 Difference 898.67 Null Hypothesis: Difference = 0 Alternative Hyp. Difference > 0 95% CI for Assumption T DF P Difference Equal Variance 2.59 4 0.0305 (−66.459, 1863.8) Unequal Variance 2.59 2.5 0.0480 (−327.29, 2124.6) Test of Equality of Variance F Num DF Den DF P 7.16 2 2 0.1226 Cases included: 6 Missing cases: 0

E. Further Embodiments of the Invention

When the term bean line EX 15340804 is used in the context of the present invention, this also includes plants modified to include at least a first desired heritable trait. Such plants may, in one embodiment, be developed by a plant breeding technique called backcrossing, wherein essentially all of the desired morphological and physiological characteristics of a variety are recovered in addition to a genetic locus transferred into the plant via the backcrossing technique. The term single locus converted plant as used herein refers to those bean plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the desired morphological and physiological characteristics of a variety are recovered in addition to the single locus transferred into the variety via the backcrossing technique.

Backcrossing methods can be used with the present invention to improve or introduce a characteristic into the present variety. The parental bean plant which contributes the locus for the desired characteristic is termed the nonrecurrent or donor parent. This terminology refers to the fact that the nonrecurrent parent is used one time in the backcross protocol and therefore does not recur. The parental bean plant to which the locus or loci from the nonrecurrent parent are transferred is known as the recurrent parent as it is used for several rounds in the backcrossing protocol.

In a typical backcross protocol, the original variety of interest (recurrent parent) is crossed to a second variety (nonrecurrent parent) that carries the single locus of interest to be transferred. The resulting progeny from this cross are then crossed again to the recurrent parent and the process is repeated until a bean plant is obtained wherein essentially all of the desired morphological and physiological characteristics of the recurrent parent are recovered in the converted plant, in addition to the single transferred locus from the nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for a successful backcrossing procedure. The goal of a backcross protocol is to alter or substitute a single trait or characteristic in the original variety. To accomplish this, a single locus of the recurrent variety is modified or substituted with the desired locus from the nonrecurrent parent, while retaining essentially all of the rest of the desired genetic, and therefore the desired physiological and morphological constitution of the original variety. The choice of the particular nonrecurrent parent will depend on the purpose of the backcross; one of the major purposes is to add some commercially desirable trait to the plant. The exact backcrossing protocol will depend on the characteristic or trait being altered to determine an appropriate testing protocol. Although backcrossing methods are simplified when the characteristic being transferred is a dominant allele, a recessive allele may also be transferred. In this instance it may be necessary to introduce a test of the progeny to determine if the desired characteristic has been successfully transferred.

In one embodiment, progeny bean plants of a backcross in which EX 15340804 is the recurrent parent comprise (i) the desired trait from the non-recurrent parent and (ii) all of the physiological and morphological characteristics of bean line EX 15340804 as determined at the 5% significance level when grown in the same environmental conditions.

Bean varieties can also be developed from more than two parents. The technique, known as modified backcrossing, uses different recurrent parents during the backcrossing. Modified backcrossing may be used to replace the original recurrent parent with a variety having certain more desirable characteristics or multiple parents may be used to obtain different desirable characteristics from each.

Many single locus traits have been identified that are not regularly selected for in the development of a new inbred but that can be improved by backcrossing techniques. Single locus traits may or may not be transgenic; examples of these traits include, but are not limited to, male sterility, herbicide resistance, resistance to bacterial, fungal, or viral disease, insect resistance, restoration of male fertility, modified fatty acid or carbohydrate metabolism, and enhanced nutritional quality. These comprise genes generally inherited through the nucleus.

Direct selection may be applied where the single locus acts as a dominant trait. An example of a dominant trait is the anthracnose resistance trait. For this selection process, the progeny of the initial cross are sprayed with anthracnose spores prior to the backcrossing. The spraying eliminates any plants which do not have the desired anthracnose resistance characteristic, and only those plants which have the anthracnose resistance gene are used in the subsequent backcross. This process is then repeated for all additional backcross generations.

Selection of bean plants for breeding is not necessarily dependent on the phenotype of a plant and instead can be based on genetic investigations. For example, one can utilize a suitable genetic marker which is closely genetically linked to a trait of interest. One of these markers can be used to identify the presence or absence of a trait in the offspring of a particular cross, and can be used in selection of progeny for continued breeding. This technique is commonly referred to as marker assisted selection. Any other type of genetic marker or other assay which is able to identify the relative presence or absence of a trait of interest in a plant can also be useful for breeding purposes. Procedures for marker assisted selection applicable to the breeding of bean are well known in the art. Such methods will be of particular utility in the case of recessive traits and variable phenotypes, or where conventional assays may be more expensive, time consuming or otherwise disadvantageous. Types of genetic markers which could be used in accordance with the invention include, but are not necessarily limited to, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., 1998).

F. Plants Derived From Bean Line EX 15340804 by Genetic Engineering

Many useful traits that can be introduced by backcrossing, as well as directly into a plant, are those which are introduced by genetic transformation techniques. Genetic transformation may therefore be used to insert a selected transgene into the bean line of the invention or may, alternatively, be used for the preparation of transgenes which can be introduced by backcrossing. Methods for the transformation of plants, including bean, are well known to those of skill in the art. Techniques which may be employed for the genetic transformation of bean include, but are not limited to, electroporation, microprojectile bombardment, Agrobacterium-mediated transformation and direct DNA uptake by protoplasts.

As is well known in the art, tissue culture of bean can be used for the in vitro regeneration of a bean plant. Tissue culture of various tissues of beans and regeneration of plants therefrom is well known. For example, reference may be had to McClean and Grafton (1989); Mergeai and Baudoin (1990); Vanderwesthuizen and Groenewald (1990); Benedicic et al. (1990); Malik and Saxena (1991).

To effect transformation by electroporation, one may employ either friable tissues, such as a suspension culture of cells or embryogenic callus or alternatively one may transform immature embryos or other organized tissue directly. In this technique, one would partially degrade the cell walls of the chosen cells by exposing them to pectin-degrading enzymes (pectolyases) or mechanically wound tissues in a controlled manner.

A particularly efficient method for delivering transforming DNA segments to plant cells is microprojectile bombardment. In this method, particles are coated with nucleic acids and delivered into cells by a propelling force. Exemplary particles include those comprised of tungsten, platinum, and preferably, gold. For the bombardment, cells in suspension are concentrated on filters or solid culture medium. Alternatively, immature embryos or other target cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an appropriate distance below the macroprojectile stopping plate.

An illustrative embodiment of a method for delivering DNA into plant cells by acceleration is the Biolistics Particle Delivery System, which can be used to propel particles coated with DNA or cells through a screen, such as a stainless steel or Nytex screen, onto a surface covered with target bean cells. The screen disperses the particles so that they are not delivered to the recipient cells in large aggregates. It is believed that a screen intervening between the projectile apparatus and the cells to be bombarded reduces the size of projectiles aggregate and may contribute to a higher frequency of transformation by reducing the damage inflicted on the recipient cells by projectiles that are too large.

Microprojectile bombardment techniques are widely applicable, and may be used to transform virtually any plant species. For example, Russell et al. (1993) disclose a method of transforming bean using electric-discharge mediated particle acceleration.

Agrobacterium-mediated transfer is another widely applicable system for introducing gene loci into plant cells. An advantage of the technique is that DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast. Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium, allowing for convenient manipulations (Klee et al., 1985). Moreover, recent technological advances in vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate the construction of vectors capable of expressing various polypeptide coding genes. The vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes. Additionally, Agrobacterium containing both armed and disarmed Ti genes can be used for transformation.

In those plant strains where Agrobacterium-mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene locus transfer. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is well known in the art (Fraley et al., 1985; U.S. Pat. No. 5,563,055). Agrobacterium-mediated transformation of P. vulgaris is described in, for example, Zhang et al. (1997); McClean et al. (1991); Lewis and Bliss (1994); and Song et al. (1995).

Transformation of plant protoplasts also can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation, and combinations of these treatments (see, e.g., Potrykus et al., 1985; Omirulleh et al., 1993; Fromm et al., 1986; Uchimiya et al., 1986; Marcotte et al., 1988). Transformation of plants and expression of foreign genetic elements is exemplified in Choi et al. (1994), and Ellul et al. (2003).

A number of promoters have utility for plant gene expression for any gene of interest including but not limited to selectable markers, scoreable markers, genes for pest tolerance, disease resistance, nutritional enhancements and any other gene of agronomic interest. Examples of constitutive promoters useful for bean plant gene expression include, but are not limited to, the cauliflower mosaic virus (CaMV) P-35S promoter, which confers constitutive, high-level expression in most plant tissues (see, e.g., Odel et al., 1985), including monocots (see, e.g., Dekeyser et al., 1990; Terada and Shimamoto, 1990); a tandemly duplicated version of the CaMV 35S promoter, the enhanced 35S promoter (P-e35S) the nopaline synthase promoter (An et al., 1988), the octopine synthase promoter (Fromm et al., 1989); and the figwort mosaic virus (P-FMV) promoter as described in U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter (P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem, the cauliflower mosaic virus 19S promoter, a sugarcane bacilliform virus promoter, a commelina yellow mottle virus promoter, and other plant DNA virus promoters known to express in plant cells.

With an inducible promoter the rate of transcription increases in response to an inducing agent. Any inducible promoter can be used in the instant invention. A variety of plant gene promoters that are regulated in response to environmental, hormonal, chemical, and/or developmental signals can be used for expression of an operably linked gene in plant cells, including promoters regulated by (1) heat (Callis et al., 1988), (2) light (e.g., pea rbcS-3A promoter, Kuhlemeier et al., 1989; maize rbcS promoter, Schaffner and Sheen, 1991; or chlorophyll a/b-binding protein promoter, Simpson et al., 1985), (3) hormones, such as abscisic acid (Marcotte et al., 1989), (4) wounding (e.g., wunl, Siebertz et al., 1989); or (5) chemicals such as methyl jasmonate, salicylic acid, or Safener. It may also be advantageous to employ organ-specific promoters (e.g., Roshal et al., 1987; Schernthaner et al., 1988; Bustos et al., 1989). Exemplary organ-specific or organ-preferred promoters include, but are not limited to, a root-preferred promoter, such as that from the phaseolin gene (Sengupta-Gopalan et al., 1985); a leaf-specific and light-induced promoter such as that from cab or rubisco (Simpson et al., 1985) and Timko et al., 1985); an anther-specific promoter such as that from LAT52 (Twell et al., 1989); a pollen-specific promoter such as that from Zm13 (Guerrero et al., 1993) or a microspore-preferred promoter such as an APG promoter (Twell et al., 1993).

Transport of protein produced by transgenes to a subcellular compartment such as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall, or mitochondrion or for secretion into the apoplast, may be accomplished by means of operably linking the nucleotide sequence encoding a signal sequence to the 5′ and/or 3′ region of a gene encoding the protein of interest. Targeting sequences at the 5′ and/or 3′ end of the structural gene may determine, during protein synthesis and processing, where the encoded protein is ultimately compartmentalized. The presence of a signal sequence directs a polypeptide to either an intracellular organelle or subcellular compartment or for secretion to the apoplast. Many signal sequences are known in the art. See, for example Becker et al. (1992); Knox et al (1987); Lerner et al. (1989); Fontes et al. (1991); Matsuoka et al. (1991); Gould et al. (1989); Creissen et al. (1991); Kalderon et al. (1984); Steifel et al. (1990).

Exemplary nucleic acids which may be introduced to the bean lines of this invention include, for example, DNA sequences or genes from another species, or even genes or sequences which originate with or are present in the same species, but are incorporated into recipient cells by genetic engineering methods rather than classical reproduction or breeding techniques. However, the term “exogenous” is also intended to refer to genes that are not normally present in the cell being transformed, or perhaps simply not present in the form, structure, etc., as found in the transforming DNA segment or gene, or genes which are normally present and that one desires to express in a manner that differs from the natural expression pattern, e.g., to over-express. Thus, the term “exogenous” gene or DNA is intended to refer to any gene or DNA segment that is introduced into a recipient cell, regardless of whether a similar gene may already be present in such a cell. The type of DNA included in the exogenous DNA can include DNA which is already present in the plant cell, DNA from another plant, DNA from a different organism, or a DNA generated externally, such as a DNA sequence containing an antisense message of a gene, or a DNA sequence encoding a synthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and could potentially be introduced into a bean plant according to the invention. Non-limiting examples of particular genes and corresponding phenotypes one may choose to introduce into a bean plant include one or more genes for insect tolerance, such as a Bacillus thuringiensis (B.t.) gene, pest resistance such as genes for fungal disease control, herbicide tolerance such as genes conferring glyphosate tolerance, and genes for quality improvements such as yield, nutritional enhancements, environmental or stress tolerances, or any desirable changes in plant physiology, growth, development, morphology or plant product(s). For example, structural genes would include any gene that confers insect tolerance including but not limited to a Bacillus insect control protein gene as described in WO 99/31248, herein incorporated by reference in its entirety, U.S. Pat. No. 5,689,052, herein incorporated by reference in its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, herein incorporated by reference it their entirety. In another embodiment, the structural gene can confer tolerance to the herbicide glyphosate as conferred by genes including, but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, herein incorporated by reference in its entirety, or glyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporated by reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes by encoding a non-translatable RNA molecule that causes the targeted inhibition of expression of an endogenous gene, for example via antisense- or cosuppression-mediated mechanisms (see, for example, Bird et al., 1991). The RNA could also be a catalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desired endogenous mRNA product (see for example, Gibson and Shillito, 1997). Thus, any gene which produces a protein or mRNA which expresses a phenotype or morphology change of interest is useful for the practice of the present invention.

G. Definitions

In the description and tables herein, a number of terms are used. In order to provide a clear and consistent understanding of the specification and claims, the following definitions are provided:

A: When used in conjunction with the word “comprising” or other open language in the claims, the words “a” and “an” denote “one or more.”

Allele: Any of one or more alternative forms of a gene locus, all of which alleles relate to one trait or characteristic. In a diploid cell or organism, the two alleles of a given gene occupy corresponding loci on a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (F₁), back to one of the parents of the hybrid progeny. Backcrossing can be used to introduce one or more single locus conversions from one genetic background into another.

Bean Yield (Tons/Acre): The recovered yield in tons/acre is the yield of the bean pods at harvest versus the means of harvest (hand picked, mechanical harvest).

Broad Adaptation: A cultivar having a broad adaptability means a cultivar or selection that will perform well in different growing conditions, locations, and seasons.

Bush Form: A USDA term about the visual look of the plant. A bean plant is: Spherical (even in width and height), Wide when the bush is wider than tall, High when the bush is taller than wide, or Stem when the individual branches protrude from the shape.

Concentrated set of pods: A concentrated set of pods is said of a plant where a high percentage of all pods on a plant set and mature at the same time so as to facilitate a single harvest.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes from different plants.

Determinate plant: A determinate plant will grow to a fixed number of nodes with a terminal floral raceme on the main stem, while an indeterminate plant continues to grow and never has a terminal floral raceme on the main stem.

Diploid: A cell or organism having two sets of chromosomes.

Dry pod color: The color of dry pods can be Buckskin (a light to pale brown), Salmon (a distinct reddish color), or Green (pale to intense) depending on the expression of the gene for persistent green.

Emasculate: The removal of plant male sex organs or the inactivation of the organs with a cytoplasmic or nuclear genetic factor conferring male sterility or a chemical agent.

Enzymes: Molecules which can act as catalysts in biological reactions.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenic plants.

Field holding ability: A bean plant that has field holding ability means a plant having pods that remain smooth and retain their color along with a firm fleshy interior as the seed approaches physiological maturity.

Genotype: The genetic constitution of a cell or organism.

Garden Bean: A plant of the genus species Phaseolus vulgaris in which the immature pods are consumed as a vegetable.

Haploid: A cell or organism having one set of the two sets of chromosomes in a diploid.

Linkage: A phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission were independent.

Machine or mechanical harvest: A machine harvestable plant means a bean plant from which the pods can be removed from the plant utilizing one of several commercial mechanical harvesters in such a manner as to reduce broken pods, clusters, and plant matter from the desired pods.

Marker: A readily detectable phenotype, preferably inherited in codominant fashion (both alleles at a locus in a diploid heterozygote are readily detectable), with no environmental variance component, i.e., heritability of 1.

Maturity: A maturity under 53 days is considered early while one between 54-59 days would be considered average or medium and one of 60 or more days would be late.

Maturity Date: Plants are considered mature when the pods have reached their maximum desirable seed size and sieve size for the specific use intended. This can vary for each end user, e.g., processing at different stages of maturity would be required for different types of consumer beans such as “whole pack,” “cut” or “french style.” The number of days is calculated from a relative planting date which depends on day length, heat units and other environmental factors.

Phenotype: The detectable characteristics of a cell or organism, which characteristics are the manifestation of gene expression.

Pod Color: A USDA term where light green color is defined by the variety Provider and a dark green color by the variety Bush Blue Lake 290. Yellow is defined as color of the wax bean Goldrush.

Pod Position: The pod position is the location of the pods within the plant. The pods can be high (near the top), low (near the bottom), or medium (in the middle) of the plant, or scattered throughout the plant.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer to genetic loci that control to some degree numerically representable traits that are usually continuously distributed.

Regeneration: The development of a plant from tissue culture.

Seed development: The rate at which seeds develop as pods reach their harvest diameter. A slow seed development characteristic will give a cultivar its field holding ability, and a larger harvest window.

Sieve Size (sv): Sieve size refers to the diameter of a pod. Sieve size 1 means pods that fall through a sieve grader which culls out pod diameters of 4.76 mm through 5.76 mm. Sieve size 2 means pods that fall through a sieve grader which culls out pod diameters of 5.76 mm through 7.34 mm. Sieve size 3 means pods that fall through a sieve grader which culls out pod diameters of 7.34 mm through 8.34 mm. Sieve size 4 means pods that fall through a sieve grader which culls out pod diameters of 8.34 mm through 9.53 mm. Sieve size 5 means pods that fall through a sieve grader which culls out pod diameters of 9.53 mm through 10.72 mm. Sieve size 6 means pods that fall through a sieve grader that will cull out pod diameters of 10.72 mm or larger.

Self-pollination: The transfer of pollen from the anther to the stigma of the same plant.

Single Locus Converted (Conversion) Plant: Plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the desired morphological and physiological characteristics of a bean variety are recovered in addition to the characteristics of the single locus transferred into the variety via the backcrossing technique and/or by genetic transformation.

Substantially Equivalent: A characteristic that, when compared, does not show a statistically significant difference (e.g., p=0.05) from the mean.

Tetraploid: A cell or organism having four sets of chromosomes.

Tissue Culture: A composition comprising isolated cells of the same or a different type or a collection of such cells organized into parts of a plant.

Transgene: A genetic locus comprising a sequence which has been introduced into the genome of a bean plant by transformation.

Triploid: A cell or organism having three sets of chromosomes.

H. Deposit Information

A deposit of bean line EX 15340804, disclosed above and recited in the claims, has been made with the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, Va. 20110-2209. The date of deposit was Nov. 28, 2006. The deposit is intended to meet all of the requirements of 37 C.F.R. §1.801-1.809. The accession number for those deposited seeds of bean line EX 15340804 is ATCC Accession No. PTA-8049. The deposit will be maintained in the depository for a period of 30 years, or 5 years after the last request, or for the effective life of the patent, whichever is longer, and will be replaced if necessary during that period.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the invention, as limited only by the scope of the appended claims.

All references cited herein are hereby expressly incorporated herein by reference.

REFERENCES

The following references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein by reference:

-   U.S. Pat. No. 5,463,175 -   U.S. Pat. No. 5,500,365 -   U.S. Pat. No. 5,563,055 -   U.S. Pat. No. 5,633,435 -   U.S. Pat. No. 5,689,052 -   U.S. Pat. No. 5,880,275 -   U.S. Pat. No. 5,378,619 -   An et al., Plant Physiol., 88:547, 1988. -   Becker et al., Plant Mol. Biol., 20:49, 1992. -   Benedicic et al., Plant Cell Tissue Org. Cult., 24:199-206, 1990. -   Bird et al., Biotech. Gen. Engin. Rev., 9:207, 1991. -   Bustos et al., Plant Cell, 1:839, 1989. -   Callis et al., Plant Physiol., 88:965, 1988. -   Choi et al., Plant Cell Rep., 13: 344-348, 1994. -   Creissen et al., Plant J., 2:129, 1991. -   Dekeyser et al., Plant Cell, 2:591, 1990. -   Ellul et al., Theor. Appl. Genet., 107:462-469, 2003. -   EP 534 858 -   Fontes et al., Plant Cell, 3:483-496, 1991. -   Fraley et al., Bio/Technology, 3:629-635, 1985. -   Fromm et al., Nature, 312:791-793, 1986. -   Fromm et al., Plant Cell, 1:977, 1989. -   Gibson and Shillito, Mol. Biotech., 7:125, 1997 -   Gould et al., J. Cell. Biol., 108:1657, 1989. -   Guerrero et al., Mol. Gen. Genetics, 244:161-168, 1993. -   Kalderon et al., Cell, 39:499-509, 1984. -   Klee et al., Bio-Technology, 3(7):637-642, 1985. -   Knox et al., Plant Mol. Biol., 9:3-17, 1987. -   Kuhlemeier et al., Plant Cell, 1:471, 1989. -   Lerner et al., Plant Physiol., 91:124-129, 1989. -   Lewis and Bliss, J. American Soc. Horticul. Sci., 119:361-366, 1994. -   Malik, and Saxena, Planta, 184(1):148-150, 1991. -   Marcotte et al., Nature, 335:454, 1988. -   Marcotte et al., Plant Cell, 1:969, 1989. -   Matsuoka et al., Proc. Natl. Acad. Sci. USA, 88:834, 1991. -   McClean and Grafton, Plant Sci., 60:117-122, 1989. -   McClean et al, Plant Cell Tiss. Org. Cult., 24:131-138, 1991. -   Mergeai and Baudoin, B.I.C. Invit. Papers, 33:115-116, 1990. -   Odel et al., Nature, 313:810, 1985. -   Omirulleh et al., Plant Mol. Biol., 21(3):415-428, 1993. -   Potrykus et al., Mol Gen. Genet., 199:183-188, 1985. -   Roshal et al., EMBO J., 6:1155, 1987. -   Russell et al., Plant Cell Reports, 12(3):165-169 (1993. -   Schaffner and Sheen, Plant Cell, 3:997, 1991. -   Schernthaner et al., EMBO J., 7:1249, 1988. -   Sengupta-Gopalan et al., Proc. Natl. Acad. Sci. USA, 82:3320-3324,     1985. -   Siebertz et al., Plant Cell, 1:961, 1989. -   Simpson et al., EMBO J., 4:2723, 1985. -   Song et al., J. Plant Physiol., 146:148-154, 1995. -   Steifel et al., Plant Cell, 2:785-793, 1990. -   Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990. -   Timko et al., Nature, 318:579-582, 1985. -   Twell et al, Mol. Gen. Genetics, 217:240-245, 1989. -   Twell et al, Sex. Plant Reprod., 6:217-224, 1993. -   Uchimiya et al., Mol. Gen. Genet., 204:204, 1986. -   Vanderwesthuizen and Groenewald, S. Afr. J. Bot., 56:271-273, 1990. -   Wang et al., Science, 280:1077-1082, 1998. -   Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990. -   WO 99/31248 -   Zhang et al., J. American Soc. Horticul. Sci., 122(3):300-305, 1997. 

1. A seed of a bean line EX 15340804 (ATCC Accession Number PTA-8049).
 2. A plant of bean line EX 15340804 (ATCC Accession Number PTA-8049).
 3. A plant part of the plant of claim
 2. 4. The plant part of claim 3, wherein said part is selected from the group consisting of a leaf, pod, pollen, an ovule and a cell.
 5. A bean plant, or a part thereof, having all the physiological and morphological characteristics of the bean plant of claim
 2. 6. A tissue culture of regenerable cells of bean line EX 15340804 (ATCC Accession Number PTA-8049).
 7. The tissue culture according to claim 6, comprising cells or protoplasts from a plant part selected from the group consisting of embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, seed and stalks.
 8. A bean plant regenerated from the tissue culture of claim 6, wherein the regenerated plant expresses all of the physiological and morphological characteristics of bean line EX 15340804 (ATCC Accession Number PTA-8049).
 9. A method of producing bean seed, comprising crossing a plant of bean line EX 15340804 (ATCC Accession Number PTA-8049) with a second bean plant.
 10. The method of claim 9, wherein the plant of bean line EX 15340804 is the female parent.
 11. The method of claim 9, wherein the plant of bean line EX 15340804 is the male parent.
 12. An F1 hybrid seed produced by the method of claim
 9. 13. An F1 hybrid plant produced by growing the seed of claim
 12. 14. A method for producing a seed of a line EX 15340804-derived bean plant comprising the steps of: (a) crossing a bean plant of line EX 15340804 (ATCC Accession Number PTA-8049) with a second bean plant; and (b) allowing seed of a EX 15340804-derived bean plant to form.
 15. The method of claim 14, further comprising the steps of: (c) crossing a plant grown from said EX 15340804-derived bean seed with itself or a second bean plant to yield additional EX 15340804-derived bean seed; (d) growing said additional EX 15340804-derived bean seed of step (c) to yield additional EX 15340804-derived bean plants; and (e) repeating the crossing and growing steps of (c) and (d) to generate further EX 15340804-derived bean plants.
 16. A method of vegetatively propagating a plant of bean line EX 15340804 comprising the steps of: (a) collecting tissue capable of being propagated from a bean plant of line EX 15340804 (ATCC Accession Number PTA-8049); (b) cultivating said tissue to obtain proliferated shoots; and (c) rooting said proliferated shoots to obtain rooted plantlets.
 17. The method of claim 16, further comprising growing plants from said rooted plantlets.
 18. A method of introducing a desired trait into bean line EX 15340804 comprising: (a) crossing a plant of line EX 15340804 (ATCC Accession Number PTA-8049) with a second bean plant that comprises a desired trait to produce F1 progeny; (b) selecting an F1 progeny that comprises the desired trait; (c) crossing the selected F1 progeny with a plant of line EX 15340804 to produce backcross progeny; (d) selecting backcross progeny comprising the desired trait and the physiological and morphological characteristic of bean line EX 15340804; and (e) repeating steps (c) and (d) three or more times in succession to produce selected third or higher backcross progeny that comprise the desired trait and all of the physiological and morphological characteristics of bean line EX 15340804 when grown in the same environmental conditions.
 19. A bean plant produced by the method of claim
 18. 20. A method of producing a plant of bean line EX 15340804 (ATCC Accession Number PTA-8049) comprising an added desired trait, the method comprising introducing a transgene conferring the desired trait into a plant of bean line EX
 15340804. 21. A plant of a bean line that exhibits root rot resistance, wherein the root rot resistance is controlled by genetic means for the expression of such trait found in bean line EX 15340804 (ATCC Accession Number PTA-8049).
 22. A seed of the plant of claim
 21. 23. A method of producing food, comprising: (a) obtaining the plant of claim 2; and (b) producing food from the plant.
 24. A method of determining the genotype of a plant of bean line EX 15340804 (ATCC Accession Number PTA-8049) comprising obtaining a sample of nucleic acids from said plant and detecting in said nucleic acids a plurality of polymorphisms.
 25. The method of claim 24, further comprising storing the results of detecting the plurality of polymorphisms on a computer readable medium.
 26. A computer readable medium produced by the method of claim
 25. 